A. Field of the Invention
The present invention is related to processes or methods to obtain extracts from agave syrup containing various phytochemicals such as polyphenols, flavonoids, saponins, phytosterols, steroids, triterpens, polycosanols and other natural products having proved antioxidant properties and more particularly to a method for using an agave syrup extract as an antioxidant, food supplement or as an anti-neoplasic, anti-carcinogenic or anti-tumoral preparation for the prevention or inhibition of growing cancers or the growth of cancer cells, such as tumors or hormone-dependent or independent cancerigen cells, from breast and lymphoma.
B. Description of Related Art
According to anthropologists, Mexico is the original source of the agave plant (Agave spp). Said plant has been used since remote times as a food source, forage, medicine, as a fiber source (henequen, wild lettuce) and as a building material among other uses. In Mesoamerica humans have always considered the importance of this plant since at least 9000 years.
Indigenous people gradually selected the sweetest agaves since they cooked the plants inside earth holes having a bed of basalt rocks and stored them for future use. Mexican indigenous civilizations described 14 different kinds of agave. The Mexoxochitli agave corresponds to the American Agave species.
Some agaves were used for curing ataxia; other agaves were used to reestablish the health in women, especially after delivery and for treating articulation pains. Agave leaves were also used for preventing scorbute and for the healing of injuries. Its main use was for the elaboration of alcoholic beverages such as pulque, mezcal, bacanora and tequila, which is produced from the Agave tequilana plant.
Taxonomically, the Agave genus pertains to the Agavaceae family. There are approximately 310 species of Agave known in the American Continent, from which 272 are found in Mexico, which led to consider Mexico as the cytogenetic origin of this important plant. The main characteristic of the Agave plant is its optimum adaptation to the arid conditions of Mesoamerica and Aridamerica. The plant pertains to the monocotiledonia group and to the lilacea order, which produce big and branched roots, succulent leaves having sunken stomas and has a photosynthetic metabolism which allows the plant to adapt to low water conditions and to take advantage of the carbon dioxide more efficiently (Granados Sánchez 1994). The chromosomic number of the Americana Agave is 2n=60.
The aguamiel or the Agave sweet extract is considered as an important beverage since it has a good nutrimental value, supplies energy, and is rich in essential amino acids essential for human growth. The aguamiel is a colorless, transparent liquid having a very sweet and pleasant flavor. It has a density of 1.049 g/cc and a total sugar content of approximately 10%. More of the 90% of the sugar content comprises sucrose (Granados Sánchez, 1994).
The vegetal material, aguamiel and syrup from the different agave kinds have been used since ancestral times for treating many maladies, specially for the prevention and treatment of diabetes, skin diseases as a scarring promoter, arthritis and cardiovascular diseases. However, the possible substances and compounds involved in the above referred biologic effects of the aguamiel are not very well known.
One of the possible action mechanisms may be related to the antioxidant activity, which has not yet been measured for the aguamiel. It is only known that the raw agave has an antioxidant capacity of 12.48 μmols trolox equivalent (ET)/g and that such value is increased to 29.38 when the agave is boiled (Wu and collaborators, 2004).
There also exist several patents related to the fructooligosaccharides (FOS) or inulin extraction from the soluble dietetic fiber. Said compounds are considered to have prebiotic, anticholesterolemic and antidiabetic effects.
In addition to the inulin the sapogenins have been an extensively researched agave compound family (Yang and col., 2006, Blunden and col., 1980, Blunden and col., 1986, Sharua and Khanna, 1980, Jin, Liu and Yang, 2003). Jin, Zhang and Yang (2004), found three new steroidal saponins (agamenosides H-J) and a new agavegenin together with six known steroids present in the residue coming from the separation of the fibers of the A. American leaves which are used in the fiber industry in China.
The steroidal saponins present in the Agave have shown to have anti-inflammatory properties which can potentially help to prevent and/or treat cancer. Da Silva and collaborators (2002) demonstrated the anti-inflammatory properties of a steroidal saponin extracted from Agave attenuate. Said steroidal saponin didn't shown the hemolytic activity shown by other saponins due to the greater affinity of the steroids for cholesterol at the erythrocytes membrane. The saponins from Dracaena surculosa, which is a plant from the Agavaceae family have been tested in vitro in order to determine its activity in leukemia promyelocytic human cells (HL-60) and only three out of 9 saponins shown a slightly cytotoxic activity (Yokosuka, Mimaki y Sashida, 2000).
Additionally, there has been reported the presence of flavonoids in the agave. The agamennone was the first flavonoid reported in the agave (Parmar and col., 1992). Subsequently, there have been found other three related flavonoids (Tinto and col. 2005). However, it hasn't been proved any biologic activity by said compounds neither its anticancerigen activity. Until now, all the reported activity related to agave syrups has been attributed to the inulin or to the sapogenins.
Japanese patent JP 11049687 claims a cutaneous preparation made of extracts from different Agave plants for external use, which stimulates the retention of water and inhibit the melamine production and thus, it is useful for dermatitis, to prevent oily skin, acne and dandruff. The Aloe Vera juice has been claimed for the treatment of mastitis in domestic cattle and for many useful common use products such as shampoos, creams for the skin, etc.
Mexican patent application of Meixueiro Valverde PCT/MX2004/0000417 disclose the production of an Agave liquid extract having anticancerigen and antiviral properties (virus sincitial of the subgroup of the Moxovirus) and that may be used as a cellular and glandular reconstituent in humans as well as in domestic animals. The extract can be used for treating asthma, sinusitis, tonsillitis, mycotic infections and intestinal parasites. The inventor relates the positive effects to phytochemical compounds having a proteinic nature which are obtained from the agavacea plant fleshy leaf cuticle and salvia. The method for the obtention of the extract comprises harvesting the agave fleshy leafs having from 4 to 15 years old; boiling the fleshy leafs for a time of from 20 to 180 minutes in water at a temperature of from 70 to 100° C. (with a water-fleshy leafs relation of 2 to 4:1); grinding the boiled fleshy leafs and filtering the resulting liquid. The extract itself comprises the viscous greyish green filtered matter having a solid content of 5%. The raw extracts from two tested agaves (alone and combined) had a good anticancerigen activity in vitro. However the inventor didn't carry out any kind of analysis in order to find which phytochemical compounds may have anticancerigen and antiviral potential. Furthermore, the related art does not disclose the anticancerigen activity of the agave syrup which was found by the applicant.
The use of the agave syrup extracts provides a source of active ingredients—not used until now—, which can be prepared and administered in a suitable way in many ways, including the treatment of cancer.
Applicant discovered that several kinds of phytochemicals from agave syrup or its extracts have potential synergic effects such as inhibiting cancer cells and or providing antioxidant effects.
Therefore, applicant developed an agave syrup extract composition (Agave spp) from several Agave varieties such as A. atrovirens, A salmiana, A. lehmanni and a related method comprising the use of its components alone or combined with other known and effective nutraceutic compounds such as vitamins A, C, and E, and/or sources of selenium for inhibiting the growth of cancer cells and/or providing an antioxidant effect.
The agave syrup extract composition of the present invention inhibits the growth of cancer cells and has antioxidant effects, for example, as an active ingredient in a food supplement and/or foods, cosmetics or medicines, or it may be an active ingredient in antineoplastic, anticancerigen or anti-tumoral preparations, for treating as well as preventing cancer or inhibiting the growth of cancer cells, such as hormone dependent or independent tumors such as breast, prostate, colon or liver cancer.
The agave syrup extract composition of the present invention comprises partially purified extracts derived from agave syrup which contain saponins, phytosterols, total phenolic compounds such as polyphenols, flavonoids and tannins which have a high antioxidant activity. Said extracts may be partially or totally purified by chromatography and/or other physic and/or chemical and/or bioseparation methods and be fractionated by a fast centrifugal partition chromatography.
The extracts may be mixed with two or more carriers, excipients and/or diluents pharmaceutically or veterinary acceptable.
The invention further comprises the isolated or purified compounds obtained from agave syrup. Said compounds are useful in an isolated way or combined at several concentrations. The compounds may be synthetically obtained as well as by its modification by several chemical and/or enzymatic processes.